Forty different sample preparation methods were tested to obtain the most informative MALDI-TOF MS protein profiles of pork meat. Extraction by 25% formic acid with the assistance of zirconia-silica beads followed by defatting by methanol:chloroform mixture (1:1, v/v) and deposition by using the layer-by-layer method was determined as the optimum sample preparation protocol. The discriminatory power of the method was then examined on samples of pork meat and meat products. The method was able to discriminate between selected salami based on the production method and brand and was able to monitor the ripening process in salami. However, it was not able to differentiate between different brands of pork ham or closely located parts of pork meat. In the latter case, a more comprehensive analysis using LC-MS/MS was used to assess the differences in protein abundance and their relation to the outputs of MALDI - TOF MS profiling.

• Klíčová slova
• MALDI-TOF mass spectrometry, Meat products, Meat ripening, Pork meat, Proteins,
• MeSH
• diskriminační analýza MeSH
• masné výrobky * analýza   MeSH
• maso analýza   MeSH
• prasata MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice * metody   MeSH
• vepřové maso analýza   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH

Fish from the pike (Esox) genus are valued in gastronomy for their superior meat quality. However, they can cause allergic reactions in sensitive consumers. This work aimed to fill the gap in the detection of pike allergens using molecular-biological techniques. New, fast, and accurate loop-mediated isothermal amplification (LAMP) and real-time PCR (qPCR) assays were designed to detect pike DNA using the parvalbumin gene as a marker. LAMP was assessed by electrophoresis, SYBR green optical detection, and real-time fluorescence detection. The latter was the most sensitive, detecting as little as 0.78 ng of pike DNA; the qPCR detection limit was 0.1 ng. The LAMP analysis took 20-70 min, which is significantly faster than qPCR. The study provides reliable detection and quantification of the parvalbumin gene in both fresh and processed samples and further highlights the versatility of the use of the parvalbumin gene for the authentication of food products and consumer protection via refined allergen risk assessment that is independent of the type of tissue or food processing method used.

• Klíčová slova
• DNA, Esox, LAMP, PCR, food allergy, food fraud,
• MeSH
• alergeny * genetika   analýza   imunologie   MeSH
• biologické markery analýza   MeSH
• diagnostické techniky molekulární MeSH
• Esocidae * genetika   imunologie   MeSH
• hodnocení rizik MeSH
• kontaminace potravin analýza   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• lidé MeSH
• parvalbuminy * genetika   imunologie   analýza   MeSH
• potravinová alergie * imunologie   MeSH
• rybí proteiny genetika   imunologie   MeSH
• techniky amplifikace nukleových kyselin metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• alergeny * MeSH
• biologické markery MeSH
• parvalbuminy * MeSH
• rybí proteiny MeSH

The infection of Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the main causes of economic losses in sericulture. Thus, it is essential to establish rapid and effective method for BmNPV detection. In the present study, we have developed a recombinase-aided amplification (RAA) to amplify the BmNPV genomic DNA at 37 °C within 30 min, and achieved a rapid detection method by coupling with a lateral flow dipstick (LFD). The RAA-LFD method had a satisfactory detection limit of 6 copies/μL of recombinant plasmid pMD19-T-IE1, and BmNPV infection of silkworm can be detected 12 h post-infection. This method was highly specific for BmNPV, and without cross-reactivity to other silkworm pathogens. In contrast to conventional polymerase chain reaction (PCR), the RAA-LFD assay showed higher sensitivity, cost-saving, and especially is apt to on-site detection of BmNPV infection in the sericulture production.

• Klíčová slova
• Bombyx mori nucleopolyhedrovirus, Lateral flow dipstick, Recombinase-aided amplification,
• MeSH
• bourec * virologie   MeSH
• DNA virů genetika   MeSH
• limita detekce MeSH
• nukleopolyhedroviry * genetika   izolace a purifikace   MeSH
• rekombinasy * metabolismus   genetika   MeSH
• senzitivita a specificita MeSH
• techniky amplifikace nukleových kyselin * metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• DNA virů MeSH
• rekombinasy * MeSH

The current study assessed the performance of the fully automated RT-PCR-based Idylla™ GeneFusion Assay, which simultaneously covers the advanced non-small cell lung carcinoma (aNSCLC) actionable ALK, ROS1, RET, and MET exon 14 rearrangements, in a routine clinical setting involving 12 European clinical centers. The Idylla™ GeneFusion Assay detects fusions using fusion-specific as well as expression imbalance detection, the latter enabling detection of uncommon fusions not covered by fusion-specific assays. In total, 326 archival aNSCLC formalin-fixed paraffin-embedded (FFPE) samples were included of which 44% were resected specimen, 46% tissue biopsies, and 9% cytological specimen. With a total of 179 biomarker-positive cases (i.e., 85 ALK, 33 ROS1, 20 RET fusions and 41 MET exon 14 skipping), this is one of the largest fusion-positive datasets ever tested. The results of the Idylla™ GeneFusion Assay were compared with earlier results of routine reference technologies including fluorescence in situ hybridization, immunohistochemistry, reverse-transcription polymerase chain reaction, and next-generation sequencing, establishing a high sensitivity/specificity of 96.1%/99.6% for ALK, 96.7%/99.0% for ROS1, 100%/99.3% for RET fusion, and 92.5%/99.6% for MET exon 14 skipping, and a low failure rate (0.9%). The Idylla™ GeneFusion Assay was found to be a reliable, sensitive, and specific tool for routine detection of ALK, ROS1, RET fusions and MET exon 14 skipping. Given its short turnaround time of about 3 h, it is a time-efficient upfront screening tool in FFPE samples, supporting rapid clinical decision making. Moreover, expression-imbalance-based detection of potentially novel fusions may be easily verified with other routine technologies without delaying treatment initiation.

• Klíčová slova
• ALK fusion, MET exon 14 skipping, RET fusion, ROS1 fusion, Idylla, NSCLC,
• MeSH
• anaplastická lymfomová kináza * genetika   MeSH
• exony * genetika   MeSH
• fúzní onkogenní proteiny * genetika   MeSH
• genová přestavba MeSH
• hybridizace in situ fluorescenční metody   MeSH
• lidé MeSH
• multiplexová polymerázová řetězová reakce MeSH
• nádorové biomarkery genetika   analýza   MeSH
• nádory plic * genetika   patologie   MeSH
• nemalobuněčný karcinom plic * genetika   patologie   MeSH
• protoonkogenní proteiny c-met * genetika   MeSH
• protoonkogenní proteiny c-ret * genetika   MeSH
• protoonkogenní proteiny * genetika   MeSH
• tyrosinkinasy * genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• multicentrická studie MeSH
• Názvy látek
• ALK protein, human MeSH Prohlížeč
• anaplastická lymfomová kináza * MeSH
• fúzní onkogenní proteiny * MeSH
• MET protein, human MeSH Prohlížeč
• nádorové biomarkery MeSH
• protoonkogenní proteiny c-met * MeSH
• protoonkogenní proteiny c-ret * MeSH
• protoonkogenní proteiny * MeSH
• RET protein, human MeSH Prohlížeč
• ROS1 protein, human MeSH Prohlížeč
• tyrosinkinasy * MeSH

Salinity stress can significantly delay plant growth. It can disrupt water and nutrient uptake, reducing crop yields and poor plant health. The use of strigolactone can be an effective technique to overcome this issue. Strigolactone enhances plant growth by promoting root development and improvement in physiological attributes. The current pot study used strigolactone to amend chili under no salinity and salinity stress environments. There were four treatments, i.e., 0, 10µM strigolactone, 20µM strigolactone and 30µM strigolactone. All treatments were applied in four replications following a completely randomized design (CRD). Results showed that 20µM strigolactone caused a significant increase in chili plant height (21.07%), dry weight (33.60%), fruit length (19.24%), fruit girth (35.37%), and fruit yield (60.74%) compared to control under salinity stress. Significant enhancement in chili chlorophyll a (18.65%), chlorophyll b (43.52%), and total chlorophyll (25.09%) under salinity stress validated the effectiveness of 20µM strigolactone application as treatment over control. Furthermore, improvement in nitrogen, phosphorus, and potassium concentration in leaves confirmed the efficient functioning of 20µM strigolactone compared to other concentrations under salinity stress. The study concluded that 20µM strigolactone is recommended for mitigating salinity stress in chili plants. Growers are advised to apply 20µM strigolactone to enhance their chili production under salinity stress.

• Klíčová slova
• Antioxidant, Chili chlorophyll content, Salinity stress, Strigolactone,
• MeSH
• Capsicum * MeSH
• chlorofyl a MeSH
• heterocyklické sloučeniny tricyklické * MeSH
• kafr MeSH
• laktony MeSH
• menthol MeSH
• salinita MeSH
• solný stres MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• chlorofyl a MeSH
• GR24 strigolactone MeSH Prohlížeč
• heterocyklické sloučeniny tricyklické * MeSH
• kafr MeSH
• laktony MeSH
• menthol MeSH

The purpose of this research is to determine whether the Covid-19 pandemic has had an impact on the change of organisational culture in public high schools. Additionally, if there has been a change in organisational culture, to what extent does this change differ from the preferred type? Cameron and Quinn's OCAI questionnaire was used to determine the types of organisational culture. 453 valid responses were obtained from teachers of randomly selected public secondary schools in all regions of the Czech Republic. Pre-Covid-19, the present and preferred status were assessed. It was found that initially hierarchy culture was predominant, while currently preferences for adhocracy and market culture have increased significantly, although the hierarchy type still prevails. In the type of future, respondents will see the clan of organisational culture. The shift in each type, but also in each of its dimensions in the three periods studied, provides the researcher with a theme for deeper research into the context, and for school institutions and principals to develop strategies to support the creation of a healthy organisational culture.

• Klíčová slova
• Covid-19, Evaluation, OCAI, Organisational culture, School,
• MeSH
• COVID-19 * epidemiologie   MeSH
• hodnocení programu MeSH
• lidé MeSH
• organizační kultura MeSH
• pandemie * MeSH
• průzkumy a dotazníky MeSH
• školy MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

Health conditions contribute significantly to patients' quality of life. Healthcare infrastructure and healthcare services, including their accessibility, belong to objective factors influencing their perception of their health. The growing disparity between supply and demand for specialized inpatient facilities due to the aging population calls for new solutions, including eHealth technologies. Automatized activities could be taken over by eHealth technologies that do not require a constant presence of staff. We tested whether eHealth technical solutions reduce patients' health risks on a sample of 61 patients on the covid-19 unit in Tomas Bata hospital in Zlin. We have applied the randomized control trial to select patients for the treatment and the control groups. Moreover, we tested eHealth technologies and their help to staff in the hospital. Due to the severity of the covid-19 disease and its rapid course and the size of the sample in our research, we did not demonstrate a statistically significant impact of eHealth technologies on patient health. The evaluation results confirm that even the limited number of technologies deployed proves to be an effective help for staff in critical situations like the pandemic. The main issue is psychological support to staff in hospitals and relieving stressful work.

• Klíčová slova
• Covid-19, EHealth, Impact on health, RCT,
• MeSH
• COVID-19 * MeSH
• hodnocení programu MeSH
• kvalita života MeSH
• lidé MeSH
• nemocnice MeSH
• senioři MeSH
• telemedicína * MeSH
• Check Tag
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH

Genetic and non-genetic factors contribute to breast cancer development. An epigenome-based signature capturing these components in easily accessible samples could identify women at risk. Here, we analyse the DNA methylome in 2,818 cervical, 357 and 227 matched buccal and blood samples respectively, and 42 breast tissue samples from women with and without breast cancer. Utilising cervical liquid-based cytology samples, we develop the DNA methylation-based Women's risk IDentification for Breast Cancer index (WID-BC-index) that identifies women with breast cancer with an AUROC (Area Under the Receiver Operator Characteristic) of 0.84 (95% CI: 0.80-0.88) and 0.81 (95% CI: 0.76-0.86) in internal and external validation sets, respectively. CpGs at progesterone receptor binding sites hypomethylated in normal breast tissue of women with breast cancer or in BRCA mutation carriers are also hypomethylated in cervical samples of women with poor prognostic breast cancer. Our data indicate that a systemic epigenetic programming defect is highly prevalent in women who develop breast cancer. Further studies validating the WID-BC-index may enable clinical implementation for monitoring breast cancer risk.

• MeSH
• cervix uteri cytologie   metabolismus   MeSH
• CpG ostrůvky MeSH
• epigenom MeSH
• epigenomika metody   MeSH
• epitelové buňky metabolismus   MeSH
• lidé MeSH
• metylace DNA * MeSH
• mutace MeSH
• nádorové biomarkery genetika   metabolismus   MeSH
• nádory prsu genetika   metabolismus   MeSH
• prognóza MeSH
• prsy cytologie   metabolismus   MeSH
• ROC křivka MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• nádorové biomarkery MeSH

Reverse transcription PCR (RT-PCR) is a popular method for detecting RNA viruses in plants. RT-PCR is usually performed in a classical two-step procedure: in the first step, cDNA is synthesized by reverse transcriptase (RT), followed by PCR amplification by a thermostable polymerase in a separate tube in the second step. However, one-step kits containing multiple enzymes optimized for RT and PCR amplification in a single tube can also be used. Here, we describe an RT-PCR single-enzyme assay based on an RTX DNA polymerase that has both RT and polymerase activities. The expression plasmid pET_RTX_(exo-) was transferred to various E. coli genotypes that either compensated for codon bias (Rosetta-gami 2) or contained additional chaperones to promote solubility (BL21 (DE3) with plasmids pKJE8 or pTf2). The RTX enzyme was then purified and used for the RT-PCR assay. Several purified plant viruses (TMV, PVX, and PVY) were used to determine the efficiency of the assay compared to a commercial one-step RT-PCR kit. The RT-PCR assay with the RTX enzyme was validated for the detection of viruses from different genera using both total RNA and crude sap from infected plants. The detection endpoint of RTX-PCR for purified TMV was estimated to be approximately 0.01 pg of the whole virus per 25 µL reaction, corresponding to 6 virus particles/µL. Interestingly, the endpoint for detection of TMV from crude sap was also 0.01 pg per reaction in simulated crude plant extracts. The longest RNA fragment that could be amplified in a one-tube arrangement was 2379 bp long. The longest DNA fragment that could be amplified during a 10s extension was 6899 bp long. In total, we were able to detect 13 viruses from 11 genera using RTX-PCR. For each virus, two to three specific fragments were amplified. The RT-PCR assay using the RTX enzyme described here is a very robust, inexpensive, rapid, easy to perform, and sensitive single-enzyme assay for the detection of plant viruses.

• Klíčová slova
• Capillovirus, Foveavirus, Luteovirus, Nepovirus, Polerovirus, Potexvirus, Potyvirus, RTX-PCR, Tobamovirus, Trichovirus, Tritimovirus, one-step RT-PCR, virus detection,
• MeSH
• DNA-dependentní DNA-polymerasy metabolismus   MeSH
• fylogeneze MeSH
• nemoci rostlin virologie   MeSH
• polymerázová řetězová reakce přístrojové vybavení   metody   MeSH
• RNA-viry klasifikace   genetika   izolace a purifikace   MeSH
• rostlinné viry klasifikace   genetika   izolace a purifikace   MeSH
• senzitivita a specificita MeSH
• zemědělské plodiny virologie   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA-dependentní DNA-polymerasy MeSH

The problems of the environment and human health related to the use of synthetic and broad-spectrum insecticides have increasingly motivated scientific research on different alternatives and among these, the use of green systems, such as essential oils, have been explored. Several species of the Apiaceae and Asteraceae families, aromatic herbs rich in secondary bioactive metabolites, are used in the industrial field for pharmaceutical, cosmetic, and food purposes. Different essential oils extracted from some species of these families have shown acute toxicity and attractive and/or repellent effects towards different insects. In our work, we investigated the toxic potential of Calendula incana subsp. maritima and Laserpitium siler subsp. siculum essential oils against four insect species, Sitophilus oryzae, Lasioderma serricorne, Necrobia rufipes, and Rhyzoperta dominica, which are common pests of stored products. The composition of both oils, extracted by hydrodistillation from the aerial parts of the two plants, was evaluated by GC×GC-MS. Calendula incana subsp. maritima essential oil was rich in oxygenated sesquiterpenoids, such as cubebol (35.39%), 4-epi-cubebol (22.99%), and cubenol (12.77%), while the Laserpitium siler subsp. siculum essential oil was composed mainly of monoterpene hydrocarbons, such as β-phellandrene (42.16%), limonene (23.87%), and β-terpinene (11.80%). The toxicity Petri dish bioassays indicated that C. maritima oil killed a mean of 65.50% of S. oryzae and 44.00% of R. dominica adults, indicating a higher biocidal activity in comparison with L. siculum oil, while toward the other species, no significant differences in mortality were recorded. Calendula maritima oil could be, then, considered a promising candidate for further tests as an alternative biocide toward S. oryzae and R. dominica. The possibility that the relatively high content of oxygenated sesquiterpenoids in C. maritima essential oil determines its higher biocidal activity is discussed.

• Klíčová slova
• GC×GC-MS analysis, Lasioderma serricorne, Necrobia rufipes, Rhyzopertha dominica, Sitophilus oryzae, cubebene derivatives,
• MeSH
• Apiaceae chemie   MeSH
• Asteraceae chemie   MeSH
• brouci účinky léků   MeSH
• insekticidy analýza   farmakologie   MeSH
• nosatcovití účinky léků   MeSH
• oleje prchavé analýza   farmakologie   MeSH
• oleje rostlin analýza   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• Názvy látek
• insekticidy MeSH
• oleje prchavé MeSH
• oleje rostlin MeSH